PpHgl10A (PpPel10a)

Endo-Hgl0311

(EC. 4.2.2.2) Exo-pectate lyase

CAZy Family: PL10

PROPERTIES

1. ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~45 kDa)

Figure 1. SDS-PAGE analysis of recombinant PpPel10a. M, molecular weight markers; lane 1, supernatant of lysed recombinant E. coli BL21 (DE3) cells carrying plasmid pET28a-pppel10a; lane 2, PpPel10a purified by Ni-NTA column chromatography.

2. SPECIFIC ACTIVITY

290 U/mg protein (on PGA) at pH 9.0 and 37°C.

One unit (U) of activity was defined as the amount of enzyme needed to produce 1 umol/min unsaturated oligogalacturonide

3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Figure 2. High-performance anion exchange chromatography analysis of the lytic products of citrus pectin produced by PpPel10a. Black line, citrus pectin; red line, lytic products of citrus pectin.

Figure 3. Ramie fibers treated with buffer (50 mM sodium glycine buffer, pH 9.0, negative control) (A,F); chemically (0.5% w/v sodium hydroxide (B,G), or 1% w/v sodium hydroxide (C,H)); enzymatically (1 U/mL PpPel10a) (D,I); or with enzyme-chemical combination (1 U/mL PpPel10a and 0.5% w/v sodium hydroxide) (E,J). Images A–E: extrinsic features of ramie fibers; images F–J: scanning electron microscopy images of single fibers (5000×)

4. PHYSICOCHEMICAL PROPERTIES

pH Optima: 9.0

pH Stability: 6.0-10.0

Temperature Optima: 45°C

5. STORAGE CONDITIONS

The enzyme should be stored at -20°C. For assay, this enzyme should be diluted in acetate buffer (20 mM) pH 4.0. Swirl to mix the enzyme immediately prior to use.

6. REFERENCES

[1] Zhao Y, Yuan Y, Gao J, et al. Screening of a Novel Polysaccharide Lyase Family 10 Pectate Lyase from Paenibacillus polymyxa KF-1: Cloning, Expression and Characterization[J]. Molecules. 2018, 23, 2774.